Recent advances in gene editing technologies have provided opportunities for precise modification of the genome in many types of organisms, including plants and animals. In particular, technologies based on genome editing proteins, such as zinc finger nucleases, TALENs, and CRISPR-Cas9 systems are advancing rapidly and it is now possible to target genetic changes to specific DNA sequences in the genome (see e.g., Segal, et at. (2013) Annu. Rev. Genomics Hum. Genet. 14, 135-158; Sander, et at. (2014) Nature Biotech. 32, 347-355). These can be either mutations resulting in gene knockouts or substitutions of one allele for another. Highly efficient CRISPR-mediated precise genome modification has now been demonstrated in several plants. In most plants, it is necessary to go through tissue culture to obtain modifications of the germline. Tissue culture can be mutagenic and therefore there is the likelihood of introducing unpredictable and undesired background, off-target mutations in addition to the desired changes. Also, it remains challenging to select for cells with the targeted genome modification.
What is desired is a method that does not require going through tissue culture and allows the selection of modified cells without the use of selectable transgenes. The present invention provides these and other advantages.